LOCUS       sequence-44159-e        3729 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  synthetic circular DNA
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 3729)
  AUTHORS   Najdi R, Proffitt K, Sprowl S, Kaur S, Yu J, Covey TM, Virshup DM, 
            Waterman ML
  TITLE     A uniform human Wnt expression library reveals a shared secretory 
            pathway and unique signaling activities.
  JOURNAL   Differentiation. 2012 Sep;84(2):203-13. doi: 
            10.1016/j.diff.2012.06.004. Epub 2012 Jul 9.
  PUBMED    22784633
REFERENCE   2  (bases 1 to 3729)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported Dec 15, 2025 from SnapGene Server 8.0.1
            https://www.snapgene.com
COMMENT     SGRef: number: 1; type: "Journal Article"; doi: 
            "10.1016/j.diff.2012.06.004"; journalName: "Differentiation"; date: 
            "2012-09"; volume: "84"; issue: "2"; pages: "203-13"
COMMENT     Sequence Label: Wnt5A Non-STOP
FEATURES             Location/Qualifiers
     source          1..3729
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     92..109
                     /label=L4440
                     /note="L4440 vector, forward primer"
     terminator      complement(268..295)
                     /label=rrnB T2 terminator
                     /note="transcription terminator T2 from the E. coli rrnB 
                     gene"
     terminator      complement(387..473)
                     /gene="Escherichia coli rrnB"
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB 
                     gene"
     primer_bind     522..544
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     536..553
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     537..553
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     protein_bind    569..668
                     /gene="mutant version of attL"
                     /label=attL1
                     /bound_moiety="LR Clonase(TM)"
                     /note="recombination site for the Gateway(R) LR reaction"
     protein_bind    complement(1854..1953)
                     /gene="mutant version of attL"
                     /label=attL2
                     /bound_moiety="LR Clonase(TM)"
                     /note="recombination site for the Gateway(R) LR reaction"
     primer_bind     complement(1970..1989)
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        complement(1971..1989)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(1994..2010)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(1994..2010)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(2112..2131)
                     /label=pENTR-R
                     /note="pENTR vectors, reverse primer"
     CDS             2123..2932
                     /codon_start=1
                     /gene="aph(3')-Ia"
                     /product="aminoglycoside phosphotransferase"
                     /label=KanR
                     /note="confers resistance to kanamycin in bacteria or G418 
                     (Geneticin(R)) in eukaryotes"
                     /translation="MSHIQRETSRPRLNSNMDADLYGYKWARDNVGQSGATIYRLYGKP
                     DAPELFLKHGKGSVANDVTDEMVRLNWLTEFMPLPTIKHFIRTPDDAWLLTTAIPGKTA
                     FQVLEEYPDSGENIVDALAVFLRRLHSIPVCNCPFNSDRVFRLAQAQSRMNNGLVDASD
                     FDDERNGWPVEQVWKEMHKLLPFSPDSVVTHGDFSLDNLIFDEGKLIGCIDVGRVGIAD
                     RYQDLAILWNCLGEFSPSLQKRLFQKYGIDNPDMNKLQFHLMLDEFF"
     primer_bind     complement(2190..2209)
                     /label=Kan-R
                     /note="Kanamycin resistance gene, reverse primer"
     rep_origin      3079..3667
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     3568..3587
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
ORIGIN
        1 ctttcctgcg ttatcccctg attctgtgga taaccgtatt accgcctttg agtgagctga
       61 taccgctcgc cgcagccgaa cgaccgagcg cagcgagtca gtgagcgagg aagcggaaga
      121 gcgcccaata cgcaaaccgc ctctccccgc gcgttggccg attcattaat gcagctggca
      181 cgacaggttt cccgactgga aagcgggcag tgagcgcaac gcaattaata cgcgtaccgc
      241 tagccaggaa gagtttgtag aaacgcaaaa aggccatccg tcaggatggc cttctgctta
      301 gtttgatgcc tggcagttta tggcgggcgt cctgcccgcc accctccggg ccgttgcttc
      361 acaacgttca aatccgctcc cggcggattt gtcctactca ggagagcgtt caccgacaaa
      421 caacagataa aacgaaaggc ccagtcttcc gactgagcct ttcgttttat ttgatgcctg
      481 gcagttccct actctcgcgt taacgctagc atggatgttt tcccagtcac gacgttgtaa
      541 aacgacggcc agtcttaagc tcgggcccca aataatgatt ttattttgac tgatagtgac
      601 ctgttcgttg caacaaattg atgagcaatg cttttttata atgccaactt tgtacaaaaa
      661 agcaggctcc gcggccgccc ccttcaccat gaagaagtcc attggaatat taagcccagg
      721 agttgctttg gggatggctg gaagtgcaat gtcttccaag ttcttcctag tggctttggc
      781 catatttttc tccttcgccc aggttgtaat tgaagccaat tcttggtggt cgctaggtat
      841 gaataaccct gttcagatgt cagaagtata tattatagga gcacagcctc tctgcagcca
      901 actggcagga ctttctcaag gacagaagaa actgtgccac ttgtatcagg accacatgca
      961 gtacatcgga gaaggcgcga agacaggcat caaagaatgc cagtatcaat tccgacatcg
     1021 aaggtggaac tgcagcactg tggataacac ctctgttttt ggcagggtga tgcagatagg
     1081 cagccgcgag acggccttca catacgcggt gagcgcagca ggggtggtga acgccatgag
     1141 ccgggcgtgc cgcgagggcg agctgtccac ctgcggctgc agccgcgccg cgcgccccaa
     1201 ggacctgccg cgggactggc tctggggcgg ctgcggcgac aacatcgact atggctaccg
     1261 ctttgccaag gagttcgtgg acgcccgcga gcgggagcgc atccacgcca agggctccta
     1321 cgagagtgct cgcatcctca tgaacctgca caacaacgag gccggccgca ggacggtgta
     1381 caacctggct gatgtggcct gcaagtgcca tggggtgtcc ggctcatgta gcctgaagac
     1441 atgctggctg cagctggcag acttccgcaa ggtgggtgat gccctgaagg agaagtacga
     1501 cagcgcggcg gccatgcggc tcaacagccg gggcaagttg gtacaggtca acagccgctt
     1561 caactcgccc accacacaag acctggtcta catcgacccc agccctgact actgcgtgcg
     1621 caatgagagc accggctcgc tgggcacgca gggccgcctg tgcaacaaga cgtcggaggg
     1681 catggatggc tgcgagctca tgtgctgcgg ccgtggctac gaccagttca agaccgtgca
     1741 gacggagcgc tgccactgca agttccactg gtgctgctac gtcaagtgca agaagtgcac
     1801 ggagatcgtg gaccagtttg tgtgcaagtg cattctgaag ggtgggcgcg ccgacccagc
     1861 tttcttgtac aaagttggca ttataagaaa gcattgctta tcaatttgtt gcaacgaaca
     1921 ggtcactatc agtcaaaata aaatcattat ttgccatcca gctgatatcc cctatagtga
     1981 gtcgtattac atggtcatag ctgtttcctg gcagctctgg cccgtgtctc aaaatctctg
     2041 atgttacatt gcacaagata aaaatatatc atcatgaaca ataaaactgt ctgcttacat
     2101 aaacagtaat acaaggggtg ttatgagcca tattcaacgg gaaacgtcga ggccgcgatt
     2161 aaattccaac atggatgctg atttatatgg gtataaatgg gctcgcgata atgtcgggca
     2221 atcaggtgcg acaatctatc gcttgtatgg gaagcccgat gcgccagagt tgtttctgaa
     2281 acatggcaaa ggtagcgttg ccaatgatgt tacagatgag atggtcagac taaactggct
     2341 gacggaattt atgcctcttc cgaccatcaa gcattttatc cgtactcctg atgatgcatg
     2401 gttactcacc actgcgatcc ccggaaaaac agcattccag gtattagaag aatatcctga
     2461 ttcaggtgaa aatattgttg atgcgctggc agtgttcctg cgccggttgc attcgattcc
     2521 tgtttgtaat tgtcctttta acagcgatcg cgtatttcgt ctcgctcagg cgcaatcacg
     2581 aatgaataac ggtttggttg atgcgagtga ttttgatgac gagcgtaatg gctggcctgt
     2641 tgaacaagtc tggaaagaaa tgcataaact tttgccattc tcaccggatt cagtcgtcac
     2701 tcatggtgat ttctcacttg ataaccttat ttttgacgag gggaaattaa taggttgtat
     2761 tgatgttgga cgagtcggaa tcgcagaccg ataccaggat cttgccatcc tatggaactg
     2821 cctcggtgag ttttctcctt cattacagaa acggcttttt caaaaatatg gtattgataa
     2881 tcctgatatg aataaattgc agtttcattt gatgctcgat gagtttttct aatcagaatt
     2941 ggttaattgg ttgtaacact ggcagagcat tacgctgact tgacgggacg gcgcaagctc
     3001 atgaccaaaa tcccttaacg tgagttacgc gtcgttccac tgagcgtcag accccgtaga
     3061 aaagatcaaa ggatcttctt gagatccttt ttttctgcgc gtaatctgct gcttgcaaac
     3121 aaaaaaacca ccgctaccag cggtggtttg tttgccggat caagagctac caactctttt
     3181 tccgaaggta actggcttca gcagagcgca gataccaaat actgtccttc tagtgtagcc
     3241 gtagttaggc caccacttca agaactctgt agcaccgcct acatacctcg ctctgctaat
     3301 cctgttacca gtggctgctg ccagtggcga taagtcgtgt cttaccgggt tggactcaag
     3361 acgatagtta ccggataagg cgcagcggtc gggctgaacg gggggttcgt gcacacagcc
     3421 cagcttggag cgaacgacct acaccgaact gagataccta cagcgtgagc attgagaaag
     3481 cgccacgctt cccgaaggga gaaaggcgga caggtatccg gtaagcggca gggtcggaac
     3541 aggagagcgc acgagggagc ttccaggggg aaacgcctgg tatctttata gtcctgtcgg
     3601 gtttcgccac ctctgacttg agcgtcgatt tttgtgatgc tcgtcagggg ggcggagcct
     3661 atggaaaaac gccagcaacg cggccttttt acggttcctg gccttttgct ggccttttgc
     3721 tcacatgtt
//
